Skip to main content
SpringerLink
Log in

The Effect of Fixation and Histological Preparation on Coronary Artery Dimensions

  • Published:
Annals of Biomedical Engineering Aims and scope Submit manuscript

Abstract

Histometric data are necessary for quantifying the cellular and tissue structure which serves as a basis for the construction of mathematical and integrative models. However, fixation and histological processing of tissues such as dehydration, embedding, sectioning and staining can cause dimensional changes and tissue distortion. The present study was designed to analyze the effects of one widely used fixation and histological preparation protocol for muscle tissue on several morphometric parameters of the coronary arteries. A total of 110 left anterior descending (LAD) artery segments obtained from eight pig hearts were fixed in 6.25% glutaraldehyde and embedded in JB-4 solution. The dimensions of the fixed rings in the loaded and no-load states and histological sections were compared with those of the fresh tissue using Bland–Altman scatter diagrams; i.e., the percent differences between measurements in two different states of various morphometric parameters (inner and outer circumference and wall thickness and area) against their means. We found that vascular elastin cannot be fixed, as seen by the retraction of the vessel dimensions when the loading of the fixed ring was removed. This retraction is time dependent and can lead to significant differences in wall thickness. The differences in dimensions between the histological sections and the fresh tissues in the no-load state were small for the inner and outer diameter (5.6 and 5.2%, respectively) and wall thickness and area (13.4 and 13.1%, respectively). These results are important for establishing an accurate quantitative data base of histological structure that can be related to vascular function or dysfunction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Ashley, C., and N. Feder. glycol methacrylate in histopathology. A study of central necrosis of the liver using a water-miscible plastic as embedding medium. Arch. Pathol. 81:391–397, 1966.

    PubMed  Google Scholar 

  2. Bassingthwaighte, J. B., T. Yipintsoi, and R. B. Harvey. Microvasculature of the dog left ventricular myocardium. Microvasc. Res. 7:229–249, 1974.

    Article  PubMed  Google Scholar 

  3. Bennett, H. S., A. D. Wyrick, S. W. Lee, and J. H. McNeil. Science and art in preparing tissues embedded in plastic for light microscopy, with special reference to glycol methacrylate, glass knives and simple stains. Stain Technol. 51:71–97, 1976.

    PubMed  Google Scholar 

  4. Bland, J. M., and D. G. Altman. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 8:307–310, 1986.

    Google Scholar 

  5. Boyle, J. J., J. Jenkins, I. C. McKay, A. R. McPhaden, and G. Lindop. An assessment of the distortion of arteries due to sectioning in endomyocardial biopsies. J. Pathol. 181:243–246, 1997.

    Article  PubMed  Google Scholar 

  6. Dobrin, P. Effect of histologic preparation on the cross-sectional area of arterial rings. J. Surg. Res. 61:413–415, 1996.

    Article  PubMed  Google Scholar 

  7. Ferguson, S. J., J. T. Bryant, and K. Ito. Three-dimensional computational reconstruction of mixed anatomical tissues following histological preparation. Med. Eng. Phys. 21:111–117, 1999.

    Article  PubMed  Google Scholar 

  8. Fernie, J. M., A. McLean, and D. Lamb. New Method for quantitating the medial component of pulmonary arteries. Arch. Pathol. Lab. Med. 109:843–848, 1985.

    PubMed  Google Scholar 

  9. Fung, Y. C., and S. S. Sobin. The retained elasticity of elastin under fixation agents. J. Biomech. Eng. 103:121–122, 1981.

    PubMed  Google Scholar 

  10. Gerrits, P. O., R. W. Horobin, and I. Stokroos. The effects of glycol methacrylate as a dehydrating agent on the dimensional changes of liver tissue. R. Microsc. Soc. 273–280, 1992.

  11. Girerd, X., J. J. Mourad, C. Acar, D. Heudes, S. Chiche, P. Bruneval, J. P. Mignot, E. Billaud, M. Safar, and S. Laurent. Noninvasive measurement of medium-sized artery intima-media thickness in humans: In vitro validation. J. Vasc. Res. 31:114–120, 1994.

    PubMed  Google Scholar 

  12. Guo, X., and G. S. Kassab. Distribution of stress and strain along the porcine aorta and coronary arterial tree. Am. J. Physiol. (Heart Circ. Physiol.) 286(6):H2361–H2368, 2004.

    Article  Google Scholar 

  13. Hirsch, E. Z., G. M. Chisolm, and A. Gibbons. Quantitative assessment of changes in aortic dimensions in response to in situ perfusion fixation at physiological pressures. Atherosclerosis 38:63–74, 1981.

    Article  PubMed  Google Scholar 

  14. Hoppeler, H., P. Lüthi, H. Claassen, E.R. Weibel, and H. Howald. The ultrastructure of the normal skeletal muscle. A morphometric analysis on untrained men, women and well-trained orienteers. Pflügers Arch. 344:217–232, 1973.

    Article  PubMed  Google Scholar 

  15. Hoppeler, H., O. Mathieu, R. Krauer, H. Claassen, R. B. Armstrong, E. R. Weibel. Design of the mammalian respiratory system. VI. Distribution of mitochondria and capillaries in various muscles. Respir. Physiol. 44:87–111, 1981.

    Article  PubMed  Google Scholar 

  16. Kassab, G. S. The coronary vasculature and its reconstruction. Ann. Biomed. Eng. 28:903–915, 2000.

    Article  PubMed  Google Scholar 

  17. Kassab, G. S., C. A. Rider, N. J. Tang, and Y. C. Fung. Morphometry of pig coronary arterial trees. Am. J. Physiol. 265 (Heart Circ. Physiol. 34):H350–H365, 1993.

    Google Scholar 

  18. Kumar, K. Microstructure of human arteries. J. Anat. Soc. India 50(2):127–130, 2001.

    Google Scholar 

  19. Park, J. C., R. J. Siegel, and L. L. Demer. Effect of calcification and formalin fixation on in vitro distensibility of human femoral arteries. Am. Heart J. 344–349, 1993.

  20. Mathieu-Costello, and O. Stereology. In: Handbook of Bioengineering, edited by R. Skalak and S. Chien. New York: McGraw-Hill, 1987, pp. 35.1–35.31.

    Google Scholar 

  21. Pesonen, E., P. Martimo, and J. Rapola. Histometry of the arterial wall: A new technique with the aid of automatic data processing. Lab. Invest. 30(2):550–555, 1974.

    PubMed  Google Scholar 

  22. Poole, D. C., and O. Mathieu-Costello. Analysis of capillary geometry in rat subepicardium and subendocardium. Am. J. Physiol. 259(1 Part 2):H204–H210, 1990.

    PubMed  Google Scholar 

  23. Siegel R. J., K. Swan, G. Edwalds, and M. Fishbein. Limitations of postmortem assessment of human coronary artery size and luminal narrowing: Differential effects of tissue fixation and processing on vessels with different degrees of atherosclerosis. JACC 5(2):342–346, 1985.

    PubMed  Google Scholar 

  24. Sobin, S. S., Y. C. Fung, and H. M. Tremer. The effect of incomplete fixation of elastin on the appearance of pulmonary alveoli. J. Biomech. Eng. 104:68–71, 1982.

    PubMed  Google Scholar 

  25. Tomanek, R. J., P. J. Palmer, G. L. Peiffer, K. L. Schreiber, C. L. Eastham, and M. L. Marcus. Morphometry of canine coronary arteries, arterioles, and capillaries during hypertension and left ventricular hypertrophy. Circ Res 58(1):38–46, 1986.

    PubMed  Google Scholar 

  26. Zoumi, A. X. Lu, G. S. Kassab, and B. J. Tromberg. Selective imaging of coronary artery micro-structural components using multi-photon microscopy. Biophys. J. 87(4):2778–2786, 2004.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, University of California, 204 Rockwell Engineering Center, Irvine, California, 92697-2715

    Jenny Susana Choy & Ghassan S. Kassab

  2. Department of Medicine, University of California, San Diego, California, 92122

    Odile Mathieu-Costello PhD

Authors
  1. Jenny Susana Choy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Odile Mathieu-Costello PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ghassan S. Kassab
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Choy, J.S., Mathieu-Costello, O. & Kassab, G.S. The Effect of Fixation and Histological Preparation on Coronary Artery Dimensions. Ann Biomed Eng 33, 1027–1033 (2005). https://doi.org/10.1007/s10439-005-4854-4

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10439-005-4854-4

Keywords

Search

Navigation